Changes in the glutathione thiol-disulfide status in wheat grain by foliar sulphur fertilization: consequences for the rheological properties of dough

Storage proteins and glutathione in wheat play an important role in gluten network formation and can be modified by supplementation of nitrogen (N) and sulphur (S) in wheat plants. The glutathione thiol-disulfide status and its relationship to the molecular weight distribution wheat polymeric protein and dough rheological properties have been examined after different foliar S fertilizations (S derived from micronized elemental S and NS, a mixture of N urea and elemental S) applied at the post-anthesis stage. Changes in levels of reduced glutathione (GSH), glutathione disulfide (GSSG), polymeric protein-glutathione mixed disulfide (PPSSG) were analysed by reversed phase high performance liquid chromatography, during grain development using the wheat cultivars, Soissons and Trémie. During the grain desiccation phase, S supplementation (i) increased the GSSG/GSH ratio by 23–25% (ii) induced PPSSG accumulation, and (iii) decreased the formation of SDS-unextractable polymeric protein (UPP) and its molecular mass distribution. However, simultaneous N and S supplementation results in: (i) a decrease in PPSSG formation by 20–30% and (ii) an increase of UPP by 7–18% by enhancing both the branching of the aggregated proteins and their molecular weight. The mixograph parameters show that all forms of endogenous glutathione are linked to dough weakening and are negatively correlated with dough mixing tolerance, dough strength and consistency, while UPP is positively correlated with dough strength and consistency. These findings indicate that S nutrition influences dynamics of the glutathione forms in the grain and results in modification the degree of polymerization of storage protein. Thus both the changes in the form of glutathione and protein polymerization influence the rheological properties of dough.     

小麦远缘杂交后代节燕98-2分离类型贮藏蛋白的研究

采用A-PAGE和SDS-PAGE聚丙烯酰胺凝胶电泳方法,对远缘组合分离出来的节燕98-2类型入选11个遗传性基本稳定的具有高产、多抗的小麦株系的醇溶蛋白和高分子量谷蛋白亚基进行了分析.结果表明,在A-PAGE电泳分析中,11个供试株系具有11种不同的醇溶蛋白带型.在SDS-FAGE电泳分析中,出现了7种不同的高分子量谷蛋白亚基(HMW-GS)及6种亚基组合类型,优质亚基及亚基组合所占的比例较少,品质评分偏低,其变幅为5～8分,平均为6.36分.但在所分析的材料中,出现了一个少见的特殊亚基:2 10 12.并研究了这些HMW-GS和组合频率及特点.11个株系中7个具有45 10优质亚基和2个具有2*亚基,它们可供小麦优质育种利用.研究表明,通过远缘杂交能够选育出具有高产、抗病和优质的小麦新材料.     

高分子量麦谷蛋白亚基的品质效应研究进展

介绍了小麦高分子量谷蛋白亚基(HMW—GS)的遗传，不同基因位点及位点内各等位亚基对品质的效应等研究进展，提出了通过回交转育创建几套不同高分子量谷蛋白亚基的近等基因系群，然后在同一遗传背景和不同遗传背景条件下分析高分子量谷蛋白亚基及其组合的品质效应及与遗传基础的关系。     

小麦低分子量谷蛋白一级结构分析

用数学统计方法和蛋白质序列分析软件对部分已测序的17种低分子量走谷蛋白亚基含氮量、氨基酸含量及一级结构序列进行比较分析。以期从氨基酸层次上揭示其结构和功能及进化上的关系．结果表明：(1)其平均含氮量为17．8％;(2)氨基酸含量基本恒定。以谷氨酰胺的含量最多。且常连接在一起;(3)存在119个保守性氨基酸残基扣两个保守性片段。一个是“QQQ-PPFSQQ”。一个是“IP-VHPSILQQLNPCKVFLQQ—C—P—AM-Q-LARSQM-QS-CHVMQQQCCQQL—QIPQQSRYEAI-AI-YSIILQEQQ”(“-”表示各序列中不同的氨基酸);(4)X84960、X84961、Y14104;X13306、U86025、U86027、U86029两组亚基序列中各具有一定程度的同源性。     

陕西省小麦品种资源高分子量谷蛋白亚基组成研究

研究了陕西省主要小麦资源高分子量谷蛋白亚基组成及其与加工品质的关系。结果表明 ,农家种亚基类型单一 ,育成种和早期国外种亚基类型分布不尽合理 ,3类材料均缺乏优质亚基 ;近期国外品种 Glu- 13个基因位点优质亚基数量较多 ,特别是 5 +10亚基 ,应加强引进、研究和利用 ;亚基 1(Glu- A1a) ,14 +15 (Glu- B1h) ,17+18(Glu- B1i)和 5 +10 (Glu- D1d)分别对多种加工品质性状效应较大 ,均为优质亚基 ;3个基因位点对加工品质的贡献值大小次序为 Glu- D1>Glu- A1>Glu- B1。     

中国小麦品种资源Glu-1位点组成概况及遗传多样性分析

 分析了 5 12 9份中国小麦初选核心种质样品HMW GS的组成情况 ,其中地方品种 345 9份、育成品种(系 ) 16 70份。这些材料作为初级核心种质基本代表了保存在国家长期库中的普通小麦种质资源的遗传多样性 ,覆盖了中国小麦栽培的 10大生态区。总体来看 ,在Glu A1、Glu B1和Glu D13个位点上的主要等位变异分别为null、7+8和 2 +12。育成品种中 1、7+9、14 +15、5 +10和 5 +12亚基 (对 )的频率比地方品种有很大的提高。在Glu 1位点上 ,地方品种与育成品种的遗传丰富度差异甚微 ,但育成品种的遗传离散度指数却显著高于地方品种。在 3个位点中 ,Glu B1位点的多样性最丰富 ,其次为Glu D1位点 ,Glu A1位点的多样性最差。从生态区来讲 ,地方品种变异类型最丰富的 3个大区是黄淮冬麦区、西北春麦区和西南冬麦区 ;选育品种最丰富的 4个大区是西南冬麦区、黄淮冬麦区、长江中下游冬麦区和北部冬麦区。由于广泛的引种、杂交、选择以及亲本选配中的偏爱 ,造成许多生态区遗传离散度指数高低与遗传丰富度出现相矛盾的现象 ,这点在长江中下游冬麦区材料中表现尤为突出。育成品种与地方品种间遗传分化系数分析表明 ,现代引种和杂交育种使我国小麦品种“群体”遗传组成和结构发生了质的变化。     

Kernel vitreousness and protein content: Relationship,interaction and synergistic effects on durum wheat quality

Four sets of durum samples were used in this study to further understand the interrelationships among hard vitreous kernels (HVK), protein content, and pigment concentration, with a focus on the interaction and synergistic effects of protein content and vitreousness on durum quality. HVK level increases with higher protein content in the range of 9.5–12.5%, but this relationship is less evident in durum samples with high protein content (12.5–14.5%). Both protein content and kernel vitreousness can significantly affect durum milling quality. White starchy kernels (WSK) in low protein durum have a very detrimental impact on milling and pasta processing quality, but high protein content can mitigate the adverse impact of WSK on durum quality. Although protein content plays a dominant role, higher HVK might contribute positively to pasta firmness. There was no significant difference in yellow pigment content between HVK and WSK. However, pigment loss from semolina to dough was higher for WSK than HVK. Despite the difference in protein content, HVK and WSK have little difference in gluten strength. The monomeric protein was preferentially accumulated in HVK. The glutenin proteins of HVK and WSK were similar in the ratios of 1Bx/1By and HMW/LMW-GS.     

Disulphide structure of high-molecular-weight (HMW-) gliadins as affected by terminators

This study aimed at elucidating SS-bonds of HMW-gliadins (HGL) from wheat with the focus on terminators of glutenin polymerisation. HGL from wheat flour extracts non-treated or treated with the S-alkylation reagent N-ethylmaleinimide (NEMI) were compared. HGL from wheat flour Akteur were isolated, hydrolysed with thermolysin and the resulting peptides pre-separated by gel permeation chromatography and analysed by liquid chromatography/mass-spectrometry using alternating electron transfer dissociation/collision-induced dissociation. Altogether, 22 and 28 SS-peptides from samples without and with NEMI treatment, respectively, were identified. Twenty-six peptides included standard SS-bonds of α- and γ-gliadins, high-molecular-weight and low-molecular-weight glutenin subunits. Eleven SS-bonds were identified for the first time. Fifteen peptides unique to HGL contained cysteine residues from gliadins with an odd number of cysteines (ω5-, α- and γ-gliadins). Thus, gliadins with an odd number of cysteines, glutathione and cysteine had acted as terminators of glutenin polymerisation. Decisive differences between samples without and with NEMI treatment were not obvious showing that the termination of polymerisation was already completed in the flour. The two HGL samples, however, were different in the majority of ten peptides that included disulphide-linked low-molecular-weight (LMW) thiols such as glutathione and cysteine with the former being enriched in the non-treated HGL-sample.     

Polymerisation of gluten proteins in developing wheat grain as affected by desiccation

The breadmaking quality of wheat is affected by the composition of gluten proteins and the polymerisation of subunits that are synthesised and accumulated in developing wheat grain. The biological mechanisms and time course of these events during grain development are documented, but not widely confirmed. Therefore, the aim of this study was to monitor the accumulation of gluten protein subunits and the size distribution of protein aggregates during grain development. The effect of desiccation on the polymerisation of gluten proteins and the functional properties of gluten were also studied. The results showed that the size of glutenin polymers remained consistently low until yellow ripeness (YR), while it increased during grain desiccation after YR. Hence, this polymerisation process was presumed to be initiated by desiccation. A similar polymerisation event was also observed when premature grains were dried artificially. The composition of gluten proteins, the ratios of glutenin to gliadin and high molecular weight-glutenin subunits to low molecular weight-glutenin subunits, in premature grain after artificial desiccation showed close association with the size of glutenin polymers in artificially dried grain. Functional properties of gluten in these samples were also associated with polymer size after artificial desiccation.     

Survey and new insights in the application of PCR‐based molecular markers for identification of HMW‐GS at the Glu‐B1 locus in durum and bread wheat

Wheat, among all cereal grains, possesses unique characteristics conferred by gluten; in particular, high molecular weight glutenin subunits (HMW‐GS) are of considerable interest as they strictly relate to bread‐making quality and contribute to strengthening and stabilizing dough. Thus, the identification of allelic composition, in particular at the Glu‐B1 locus, is very important to wheat quality improvement. Several PCR‐based molecular markers to tag‐specific HMW glutenin genes encoding Bx and By subunits have been developed in recent years. This study provides a survey of the molecular markers developed for the HMW‐GS at the Glu‐B1 locus. In addition, a selection of molecular markers was tested on 31 durum and bread wheat cultivars containing the By8, By16, By9, Bx17, Bx6, Bx14 and Bx17 Glu‐B1 alleles, and a new assignation was defined for the ZSBy9_aF1/R3 molecular marker that was specific for the By20 allele. We believe the results constitute a practical guide for results that might be achieved by these molecular markers on populations and cultivars with high variability at the Glu‐B1 locus.